Experiment 2: Dilution & Beers Law

Purpose:
In this experiment, you will:
Prepare a CuSO4 standard solution.
Measure the absorbance value of each standard solution.
Find the relationship between absorbance and concentration of a solution.
Determine the concentration of an unknown CuSO4 solution.
Reference:
Modified by Dr. Gautreaux from D. Holmquist, J. Randall, D. Volz. Chemistry with Vernier, 1st
edition; Vernier Software & Technology: Beaverton, OR, 2007; 17-1 17-4.
FigureTheory:
1
Background

The primary objective of this experiment is to determine the

concentration of an unknown copper (II) sulfate solution. You will be using a Colorimeter or
Spectrometer. The wavelength of light used should be one that is absorbed by the solution. The
CuSO4 solution used in this experiment has a deep blue color, so Colorimeter users will be
instructed to use the red LED. Spectrometer users will determine an appropriate wavelength
based on the absorbance spectrum of the solution. The light striking the detector is reported as
absorbance or percent transmittance. A higher concentration of the colored solution absorbs
more light (and transmits less) than a solution of lower concentration.
You will prepare five copper sulfate solutions of known concentration (standard solutions). Each
is transferred to a small, rectangular cuvette that is placed into the Colorimeter or Spectrometer.
The amount of light that penetrates the solution and strikes the photocell is used to compute the
absorbance of each solution. When a graph of absorbance vs. concentration is plotted for the
standard solutions, a direct relationship should result, as shown in Figure 1. The direct
relationship between absorbance and concentration for a solution is known as Beers law.
You will determine the concentration of an unknown CuSO4 solution by measuring its
absorbance. By locating the absorbance of the unknown on the vertical axis of the graph, the
corresponding concentration can be found on the horizontal axis (follow the arrows in Figure 1).
The concentration of the unknown can also be found using the slope of the Beers law curve and
the equation of a straight line, y = mx + b.

Chemical Hazards:
Name
Formula/Structure
Copper II Sulfate
CuSO4

CHEM 1316L X2 BEERS LAW

Hazards
PPE
Skin and respiratory Goggles, gloves, and

irritant; moderately lab coat

toxic by ingestion
and inhalation
Pre-Lab Questions:
1. How would you prepare from the stock solution of 0.5 M CuSO 4, 5 mL of 0.1M, 0.2M,
0.3M, and 0.4M, and 0.5M CuSO4. Show your calculations in your submission
2. A calibration pot of absorbance vs. concentration was obtained with the slope of the bestfit straight line as 1220 M-1 . The absorbance of a dilute CuSO4 solution was 0.72. What
is the concentration of the dilute CuSO4 solution?
3. Dye Lou Shun was asked to prepare 10 mL of a 0.6 M solution of HCl from a 2 M stock
solution of HCl. How many mL of water and 2 M HCl are needed to prepare 10 mL of
0.6M HCl?
4. Way Boat needs to make 100mL of a 1 M solution of CoCl2. He looks in the lab cabinet
and finds CoCl2 6 H2O. Calculate the amount of CoCl2 6 H2O needed to make 100 mL
of 1M CoCl2. Write 2 3 sentences explaining how to make this solution.
5. How many mL of 0.75 M HNO3 are required to make 30 mL of 0.25 M HNO3?
Procedure:
1. Obtain and wear goggles. CAUTION: Be careful not to ingest any CuSO4 solution or spill
any on your skin. Inform your teacher immediately in the event of an accident.
2. Add about 30 mL of 0.40 M CuSO4 stock solution to a 100 mL beaker. Add about 30 mL of
distilled water to another 100 mL beaker.
3. Label four clean, dry, test tubes 14 (the fifth solution is the beaker of 0.40 M CuSO4). Fill in
the table below to determine how to make 8 mL of each CuSO4 solution listed. Once you
have determined how to make each solution, thoroughly mix each solution with a stirring rod.
Clean and dry the stirring rod between stirrings. Keep the remaining 0.40 M CuSO4 in the 100
mL beaker to use in the fifth trial.
Test tube
number
1
2
3
4
5

Volume of initial
0.40 M CuSO4
(mL)

Volume of Distilled
H2O
(mL)

Final
Concentration of
CuSO4 (M)
0.08
0.16
0.24
0.32
0.40

4. Prepare a blank by filling an empty cuvette 3/4 full with distilled water. To correctly use a
Colorimeter cuvette, remember:
All cuvettes should be wiped clean and dry on the outside with a tissue.
Handle cuvettes only by the top edge of the ribbed sides.

CHEM 1316L X2 BEERS LAW

 All solutions should be free of bubbles.

Always position the cuvette so the light passes through the clear sides.
5. Connect the Colorimeter to LabQuest and choose New from the File menu.
6. Calibrate the Colorimeter.
a. Place the blank in the cuvette slot of the Colorimeter and close the lid.
b. Press the < or > buttons on the Colorimeter to set the wavelength to 635 nm (Red). Then
calibrate by pressing the CAL button on the Colorimeter. When the LED stops flashing,
the calibration is complete.
7. Set up the data-collection mode.
a. On the Meter screen, tap Mode. Change the mode to Events with Entry.
b. Enter the Name (Concentration) and Units (mol/L). Select OK.
8. You are now ready to collect absorbance-concentration data for the five standard solutions.
a. Start data collection.
b. Empty the water from the cuvette. Using the solution in Test Tube 1, rinse the cuvette
twice with ~1 mL amounts and then fill it 3/4 full. Wipe the outside with a tissue and place
it in the device (Colorimeter or Spectrometer). Close the lid on the Colorimeter.
c. When the value displayed on the screen has stabilized, tap Keep and enter 0.080 as the
concentration in mol/L. Select OK. The absorbance and concentration values have now
been saved for the first solution.
d. Discard the cuvette contents as directed by your instructor. Using the solution in Test
Tube 2, rinse the cuvette twice with ~1 mL amounts, and then fill it 3/4 full. Place the
cuvette in the device, wait for the value displayed on the screen to stabilize, and tap Keep.
Enter 0.16 as the concentration in mol/L. Select OK.
e. Repeat the procedure for Test Tube 3 (0.24 M) and Test Tube 4 (0.32 M), as well as the
stock 0.40 M CuSO4. Note: Wait until Step 10 to test the unknown.
f. Stop data collection.
g. To examine the data pairs on the displayed graph, tap any data point. As you tap each data
point, the absorbance and concentration values are displayed to the right of the graph.
Record the absorbance and concentration data values in your data table.
9. Display a graph of absorbance vs. concentration with a linear regression curve.
a.
b.
c.
d.

Choose Graph Options from the Graph menu.

Select Autoscale from 0 and select OK.
Choose Curve Fit from the Analyze menu.
Select Linear as the Fit Equation. The linear-regression statistics for these two data
columns are displayed for the equation in the form
y = mx + b
where x is concentration, y is absorbance, a is the slope, and b is the y-intercept.
Note: One indicator of the quality of your data is the size of b. It is a very small value if
the regression line passes through or near the origin. The correlation coefficient, r,
indicates how closely the data points match up with (or fit) the regression line. A value of
1.00 indicates a nearly perfect fit.

CHEM 1316L X2 BEERS LAW

e. Select OK. The graph should indicate a direct relationship between absorbance and
concentration, a relationship known as Beers law. The regression line should closely fit
the five data points and pass through (or near) the origin of the graph.
10. Determine the absorbance value of the unknown NiSO4 solution.
a. Tap the Meter tab.
b. Obtain about 5 mL of the unknown CuSO4 in another clean, dry, test tube. Record the
number of the unknown in your data table.
c. Rinse the cuvette twice with the unknown solution and fill it about 3/4 full. Wipe the
outside of the cuvette and place it into the device.
d. Monitor the absorbance value. When this value has stabilized, record it in your data table.
11. Discard the solutions as directed by your instructor.

PROCESSING THE DATA

1. To determine the concentration of the unknown CuSO4 solution, interpolate along the
regression line to convert the absorbance value of the unknown to concentration.
a. Tap the Graph tab.
b. Choose Interpolate from the Analyze menu.
c. Tap any point on the regression curve (or use the or keys on LabQuest) to find the
absorbance value that is closest to the absorbance reading you obtained in Step 10. The
corresponding NiSO4 concentration, in mol/L, will be displayed to the right of the graph.
d. Record the concentration value in your data table.
2. In your notebook, draw a graph of absorbance vs. concentration and unknown concentration
displayed.
Post Lab Questions:
1. What is the molar concentration of your unknown sample of copper (II) sulfate solution?
2. What factors are included in the Beers law expression for determining how much light
passes through a liquid solution?
3. How would your test results be affected if you left fingerprints on the sides of the cuvette
in line with the light path of the spectrometer (or colorimeter)?
4. Could this method of testing be used to determine the concentration of a NaCl solution?
5. During your Beers Law experiment, you got the following standard curve plot. What is
incorrect about this plot, and what error most likely occurred to get a plot like this?

CHEM 1316L X2 BEERS LAW

6. Explain, in 2 3 sentences, how you would determine the concentration of an unknown

solution using a Beers Law Plot.

CHEM 1316L X2 BEERS LAW